EP0189707A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Recombinant DNA which can be introduced into plant cells
Assignee: Bayer CropScience NV
•
Filing: 1985-12-20
•
Granted: —
•
Expires: 2005-12-20
Abstract
Abstract The invention relates to chimaeric genes which encode a fusion protein consisting of a transit peptide of a chloroplast protein-precursor, linked directly or indirectly to another protein. It also relates to recombinant vectors containing said chimeric genes suitable for the transformation of plant cells, whereby said another protein can be translocated and processed in the chloroplats.
Inventors
- 👤 Luis Herrera-Estrella
- 👤 Guido Van Den Broeck
- 👤 Marc Van Montagu
- 👤 Peter Schreier
- 👤 Jeff Max Planck Inst. für Zuchtungsforsch. Schell
- 👤 Hans J. Bohnert
- 👤 Anthony R. Cashmore
- 👤 Michael P. Timko
- 👤 Albert P. Kausch
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 16:47
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/82; C12N15/8216; C12N15/8221
Claims Analysis
Total Claims: 38
Claim 1
Independent Claim
147 words
A recombinant DNA which can be introduced into determined plant cells and which contains a chimaeric gene comprising a first nucleic acid and a second nucleic acid recombined with each other, said first nucleic acid and said second nucleic acid being of different origins, wherein said first nucleic acid contains a first coding sequence which has essential sequence homology with a natural gene sequence coding for a transit peptide belonging to a precursor comprising at least the N-terminal sub-unit of a chloroplast protein capable of being transported into or processed within the chloroplast of said plant cells and wherein said second nucleic acid contains a second coding sequence distinct of the gene sequence coding for said chloroplast protein or chloroplast protein sub-unit, said second nucleic acid being located downstream of said first nucleic acid in the direction of the transcription of their first and second sequences respectively.
Claim 2
Dependent Claim
147 words
A recombinant DNA which can be introduced into determined plant cells and which contains a chimaeric gene comprising a first nucleic acid and a second nucleic acid recombined with each other, said first nucleic acid and said second nucleic acid being of different origins, wherein said first nucleic acid contains a first coding sequence which has essential sequence homology with a natural gene sequence coding for a transit peptide belonging to a precursor comprising at least the N-terminal sub-unit of a chloroplast protein capable of being transported into or processed within the chloroplast of said plant cells and wherein said second nucleic acid contains a second coding sequence distinct of the gene sequence coding for said chloroplast protein or chloroplast protein sub-unit, said second nucleic acid being located downstream of said first nucleic acid in the direction of the transcription of their first and second sequences respectively.
Claim 3
Dependent Claim
57 words
The recombinant DNA of claim 1 wherein said first nucleic acid further contains a third sequence corresponding to at least part of a nucleic acid encoding the N-terminal cytoplasmic subunit of a chloroplast protein downstream of said first sequence and wherein the extremity of said third sequence coincides substantially with the extremity of said first nucleic acid.
Claim 4
Dependent Claim
28 words
The recombinant DNA of claim 2 wherein said third sequence does not extend beyond the nucleotides encoding the C-terminal extremity of the cytoplasmic subunit of said chloroplast protein.
Claim 5
Dependent Claim
24 words
The recombinant DNA of claim 1, wherein said first coding sequence and at least part of said third sequence belong to a same exon.
Claim 6
Dependent Claim
13 words
The recombinant DNA of claim 4, wherein said third sequence comprises an intron.
Claim 7
Dependent Claim
37 words
The recombinant DNA of claim 3 wherein said third sequence is limited to the region of high homology which the N-terminal parts of cytoplasmic precursor-subunits of chloroplast proteins from soybean, pea, duck-weed and wheat have in common.
Claim 8
Dependent Claim
14 words
The recombinant DNA of claim 6 wherein said third sequence encodes the pentapeptide M-Q-V-W-P.
Claim 9
Dependent Claim
27 words
The recombinant DNA of claim 1 wherein the end of said first nucleic acid coincides with or is close to the end of said first coding sequence.
Claim 10
Dependent Claim
32 words
The recombinant DNA of claim 8, wherein said second coding sequence is substantially free of sequence homology with the N-terminal portions of gene sequences coding for a chloroplast protein or chloroplast sub-unit.
Claim 11
Dependent Claim
45 words
The recombinant DNA of claim 8, wherein the nucleotide sequence next to the first sequence coding for the transit peptide is substantially free of homology with the nucleotide sequence coding for the first aminoacids (methionine not included) of the N-terminal part of said chloroplast protein.
Claim 12
Dependent Claim
22 words
The recombinant DNA of claim 8 which is free of an intron between said first coding sequence and said second coding sequence.
Claim 13
Dependent Claim
40 words
The recombinant DNA of claim 11, wherein the first codon of said second coding sequence is a codon coding for methionine and the latter codon is adjacent to the last codon of the first sequence coding for the transit peptide.
Claim 14
Dependent Claim
62 words
The recombinant DNA of claim 1, which further comprises a promoter region upstream of said first sequence coding for the transit peptide and so positioned that both said first sequence and said second sequence are under transcriptional control of said promoter region and wherein said promoter region contains a promoter sequence which is recognized by the polymerases of said determined plant cells.
Claim 15
Dependent Claim
33 words
The recombinant DNA of claim 13, wherein the promoter region is the promoter region (homologous promoter region) normally associated with said first sequence coding for the transit peptide in the corresponding natural gene.
Claim 16
Dependent Claim
19 words
The recombinant DNA of claim 14, wherein the promoter region is foreign (heterologous promoter region) to said first sequence.
Claim 17
Dependent Claim
33 words
The recombinant DNA of claim 1, wherein said first sequence codes for a transit peptide normally associated with a gene sequence coding for the small sub-unit of ribulose - 1.5 - biphosphate carboxylase.
Claim 18
Dependent Claim
15 words
The recombinant DNA of claim 1, wherein said second sequence codes for a bacterial protein.
Claim 19
Dependent Claim
15 words
The recombinant DNA of claim 1, wherein said second sequence codes for a plant protein.
Claim 20
Dependent Claim
17 words
The recombinant of claim 18, wherein said plant protein is a chloroplast protein or a sub-unit thereof.
Claim 21
Dependent Claim
15 words
The recombinant of claim 19, wherein said second sequence codes for a mutated chloroplast protein.
Claim 22
Dependent Claim
25 words
The recombinant DNA of claim 1, which is a vector which can be introduced and maintained in said plants, said vector comprising said chimaeric gene.
Claim 23
Dependent Claim
80 words
The recombinant DNA of claim 20 which is a vector which, in addition to said chimaeric gene, also comprises a DNA fragment having essential sequence homology with the DNA fragment which in a Ti-plasmid comprises the sequences encoding the essential functions capable of causing the transfer of the T-DNA fragment of said Ti-plasmid into said plant cells, said chimaeric gene being so positioned with respect to said chimaeric gene that the latter can be transferred into said plant cells too.
Claim 24
Dependent Claim
25 words
The vector recombinant DNA of claim 22 which contains a T-DNA border sequence and wherein said chimaeric gene is positioned close to said border sequence.
Claim 25
Dependent Claim
22 words
The vector of claim 32 which comprises two border sequences and wherein said chimaeric gene is positioned between said two border sequences.
Claim 26
Dependent Claim
139 words
A process for causing and controlling the introduction of a determined protein or polypeptide into the chloroplasts of determined plant cells which comprises introducing into said plant cells a chimaeric gene comprising a first nucleic acid recombined with a distinct second nucleic acid, wherein the first nucleic acid contains a first coding sequence which has an essential sequence homology with a natural gene sequence coding for a transit peptide belonging to a precursor comprising at least the N-terminal sub-unit of a chloroplast protein capable of being transported into or processed within the chloroplast of said plant cells, and wherein said second nucleic acid contains a sequence coding for said determined protein or polypeptide, said second nucleic acid being located downstream of the first nucleic acid in the direction of the respective transcriptions of their first and second sequences respectively.
Claim 27
Dependent Claim
139 words
A process for causing and controlling the introduction of a determined protein or polypeptide into the chloroplasts of determined plant cells which comprises introducing into said plant cells a chimaeric gene comprising a first nucleic acid recombined with a distinct second nucleic acid, wherein the first nucleic acid contains a first coding sequence which has an essential sequence homology with a natural gene sequence coding for a transit peptide belonging to a precursor comprising at least the N-terminal sub-unit of a chloroplast protein capable of being transported into or processed within the chloroplast of said plant cells, and wherein said second nucleic acid contains a sequence coding for said determined protein or polypeptide, said second nucleic acid being located downstream of the first nucleic acid in the direction of the respective transcriptions of their first and second sequences respectively.
Claim 28
Dependent Claim
87 words
The process of claim 25 which comprises transforming said pant cells containing said chloroplasts with a vector which, in addition to said chimaeric gene, also comprises a DNA fragment having essential sequence homology with the DNA fragment which in a Ti-plasmid comprises the sequences encoding the essential functions capable of causing the transfer of the T-DNA fragment of said Ti-plasmid into said plant cells, said chimaeric gene being so positioned with respect to said chimaeric gene that the latter can be transferred into said plant cells too.
Claim 29
Dependent Claim
25 words
The process of claim 26 wherein said vector comprises two T-DNA border sequences and wherein said chimaeric gene is positionned between said two border sequences.
Claim 30
Dependent Claim
60 words
The process of claim 27, wherein the first nucleic acid of said chimaeric gene further contains a third sequence corresponding to at least part of a nucleic acid encoding the N-terminal cytoplasmic subunit of a chloroplast protein downstream of said first sequence and wherein the extremity of said third sequence coincides substantially with the extremity of said first nucleic acid.
Claim 31
Dependent Claim
27 words
The process of claim 28 wherein the first coding sequence of said chimaeric gene and at least part of said third sequence belong to a same exon.
Claim 32
Dependent Claim
36 words
The process of claim 29, wherein said third sequence is limited to the region of high homology which the N-terminal parts of cytoplasmic precursor-subunits of chloroplast proteins from soybean, pea, duck-weed and wheat have in common.
Claim 33
Dependent Claim
49 words
The process of claim 27 wherein the nucleotide sequence next to the first sequence of said chimaeric gene, which codes for the transit peptide is substantially free of homology with the nucleotide sequence coding for the first aminoacids (methionine not included) of the N-terminal part of said chloroplast protein.
Claim 34
Dependent Claim
67 words
The process of claim 26, wherein the chimaeric gene of said vector further comprises a promoter region upstream of said first sequence coding for the transit peptide and so positioned that both said first sequence and said second sequence are under transcriptional control of said promoter region and wherein said promoter region contains a promoter sequence which is recognized by the polymerases of said determined plant cells.
Claim 35
Dependent Claim
53 words
The cells of a determined plant which contain in their genome a chimaeric gene as defined in claim 1, wherein the second coding sequence of said chimeric gene encodes a polypeptide or protein foreign to said plant, the chloroplasts of said plant cells further containing detectable amounts of said foreign polypeptide or protein.
Claim 36
Dependent Claim
9 words
A culture of the plant cells of claim 33.
Claim 37
Dependent Claim
18 words
Plants or parts thereof including seeds containing in their genome a chimaeric gene as defined in claim 1.
Claim 38
Dependent Claim
14 words
Plant seeds containing in their genome a chimaeric gene as defined in claim 1.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.